Diversification of Drosophila Chloride Channel Gene by Multiple Posttranscriptional mRNA Modifications

Abstract : We have identified and analyzed a Drosophila melanogaster gene that encodes a chloride channel subunit (DrosGluCl‐α) previously shown to function as a glutamategated chloride channel in an in vitro expression system. Sequence analysis of several cDNAs corresponding to the gene revealed sequence diversity in their open reading frames at seven specific sites. Site‐specific A‐to‐G variations between cDNA and genomic sequences, consistent with RNA editing, were detected at five nucleotide positions. In addition, sequence variations among cDNA clones consistent with alternative splicing of mRNA were found at two different sites. In the 5′ region, two small adjacent exons, containing similar but distinct modular sequences, are alternatively incorporated into the mature mRNA. In the 3′ region, alternative splicing generates a variant encoding a protein with four additional amino acids just upstream of the fourth transmembrane domain. Combinations of RNA editing and alternative splicing can lead to extensive diversification of transcripts. These results give the first example of RNA editing in neurotransmitter‐gated chloride channel genes or of alternative splicing in a glutamate‐gated chloride channel gene of Drosophila.

[1]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[2]  A. Wolstenholme,et al.  Alternative splicing of a Caenorhabditis elegans gene produces two novel inhibitory amino acid receptor subunits with identical ligand binding domains but different ion channels. , 1997, Gene.

[3]  Jeffrey C. Hall,et al.  A Drosophila Calcium Channel α1 Subunit Gene Maps to a Genetic Locus Associated with Behavioral and Visual Defects , 1996, The Journal of Neuroscience.

[4]  D. Cully,et al.  Identification of a Drosophila melanogaster Glutamate-gated Chloride Channel Sensitive to the Antiparasitic Agent Avermectin* , 1996, The Journal of Biological Chemistry.

[5]  J. Vaughn,et al.  RNA editing in Drosophila 4f-rnp gene nuclear transcripts by multiple A-to-G conversions. , 1996, Journal of molecular biology.

[6]  P. Seeburg,et al.  Structural Requirements for RNA Editing in Glutamate Receptor Pre-mRNAs by Recombinant Double-stranded RNA Adenosine Deaminase (*) , 1996, The Journal of Biological Chemistry.

[7]  P. Seeburg.  The Role of RNA Editing in Controlling Glutamate Receptor Channel Properties , 1996, Journal of neurochemistry.

[8]  A. Karlin,et al.  Toward a structural basis for the function of nicotinic acetylcholine receptors and their cousins , 1995, Neuron.

[9]  S. Russek,et al.  From ion currents to genomic analysis: Recent advances in GABAA receptor research , 1995, Synapse.

[10]  D. Vassilatis,et al.  Cloning of an avermectin-sensitive glutamate-gated chloride channel from Caenorhabditis elegans , 1994, Nature.

[11]  R. ffrench-Constant,et al.  Expression of a Drosophila GABA receptor in a baculovirus insect cell system , 1993, FEBS letters.

[12]  R. ffrench-Constant,et al.  Drosophilaγ‐Aminobutyric Acid Receptor Gene Rdl Shows Extensive Alternative Splicing , 1993 .

[13]  B. Sakmann,et al.  Determinants of ca2+ permeability in both TM1 and TM2 of high affinity kainate receptor channels: Diversity by RNA editing , 1993, Neuron.

[14]  H. Betz,et al.  Glutamate receptors of Drosophila melanogaster: cloning of a kainate-selective subunit expressed in the central nervous system. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[15]  S. Nakanishi.  Molecular diversity of glutamate receptors and implications for brain function. , 1992, Science.

[16]  R. North,et al.  Calcium-activated potassium channels expressed from cloned complementary DNAs , 1992, Neuron.

[17]  B. Sakmann,et al.  A glutamate receptor channel with high affinity for domoate and kainate. , 1992, The EMBO journal.

[18]  L. Vyklický,et al.  Molecular cloning and development analysis of a new glutamate receptor subunit isoform in cerebellum , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[19]  B. Sakmann,et al.  Divalent ion permeability of AMPA receptor channels is dominated by the edited form of a single subunit , 1992, Neuron.

[20]  P. Seeburg,et al.  RNA editing in brain controls a determinant of ion flow in glutamate-gated channels , 1991, Cell.

[21]  H. Betz,et al.  Molecular cloning of an invertebrate glutamate receptor subunit expressed in Drosophila muscle. , 1991, Science.

[22]  R. ffrench-Constant,et al.  Molecular cloning and transformation of cyclodiene resistance in Drosophila: an invertebrate gamma-aminobutyric acid subtype A receptor locus. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[23]  S. Antonarakis,et al.  Cloning of the gamma-aminobutyric acid (GABA) rho 1 cDNA: a GABA receptor subunit highly expressed in the retina. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. Heinemann,et al.  Cloning of a novel glutamate receptor subunit, GluR5: Expression in the nervous system during development , 1990, Neuron.

[25]  B. Sakmann,et al.  Flip and flop: a cell-specific functional switch in glutamate-operated channels of the CNS. , 1990, Science.

[26]  B. Ganetzky,et al.  Molecular analysis of the para locus, a sodium channel gene in Drosophila , 1989, Cell.

[27]  O. Pongs,et al.  Shaker encodes a family of putative potassium channel proteins in the nervous system of Drosophila. , 1988, The EMBO journal.

[28]  Y. Jan,et al.  Multiple potassium–channel components are produced by alternative splicing at the Shaker locus in Drosophila , 1988, Nature.

[29]  A. Karlin,et al.  Acetylcholine receptor binding site contains a disulfide cross-link between adjacent half-cystinyl residues. , 1986, The Journal of biological chemistry.

[30]  A. Feinberg,et al.  A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. , 1983, Analytical biochemistry.

[31]  S. Heinemann,et al.  Cloned glutamate receptors. , 1994, Annual review of neuroscience.

[32]  M. Biel,et al.  Molecular basis for Ca2+ channel diversity. , 1994, Annual review of neuroscience.

[33]  P. Seeburg,et al.  Glutamate receptor channels: novel properties and new clones. , 1992, Trends in pharmacological sciences.

[34]  Y. Jan,et al.  Multiple potassium-channel components are produced by alternative splicing at the shaker locus in Drosophila , 1988, Nature.

[35]  Mary Lou Pardue,et al.  [38] Nucleic acid hybridization in cytological preparations , 1971 .